蛋白激酶C和JNK在溶血磷脂酸诱导人肺成纤维细胞MCP-1表达中的作用

doi:10.3969/j.issn.1004-616x.2017.01.006

Abstract

Abstract:

OBJECTIVE: To investigate the role of protein kinase C (PKC) and c-Jun N-terminal kinase (JNK) in modulating lysophosphatidic acid (LPA)-induced monocyte chemo-attractant protein-1 (MCP-1) expression in human fetal lung fibroblasts (HLF-1). METHODS: Cultured human lung fibroblasts (HLF-1) were incubated with LPA(0,1,3 and 10 μmol/L) for 0.5,6,12 and 24 h. ELISA was used to detect MCP-1 protein levels in the supernatants,and quantitative real-time PCR (qPCR) for MCP-1 mRNA levels in the cell lysate. In addition,cells were pre-incubated with PKC inhibitor bisindolylmaleimide I (0,0.1,1 and 10 μmol/L) or JNK inhibitor SP600125 (0,0.1,1 and 10 μmol/L) for 30 min,and then treated with LPA (10 μmol/L) for 2 or 6 h,and ELISA and qPCR assays were conducted. Cells were also pre-incubated with PKC inhibitor bisindolylmaleimide I (1 μmol/L) for 30 min,and then treated with LPA (10 μmol/L) for 30 min. C-Jun N-terminal phosphorylation levels in the cell lysate were measured by Western blot. RESULTS: LPA stimulated MCP-1 protein expression in dose- and time-dependent manners. The MCP-1 protein production in the LPA (10 μmol/L) treated group was 2.4 times higher than that in the untreated group (P < 0.01). MCP-1 protein production in the LPA (10 μmol/L) treated group for 24 h was 1 time higher than that in the untreated group (P < 0.01). LPA stimulated MCP-1 mRNA expression in a time-dependent manner. The MCP-1 mRNA expression in the LPA (10 μmol/L) treated group for 2 h was 5.3 times higher than that in the untreated group (P < 0.01). PKC inhibitors bisindolylmaleimide I and JNK SP600125 clearly inhibited LPA-induced MCP-l mRNA and protein expressions. The LPA (10 μmol/L)-induced MCP-1 protein production was reduced by 60% with bisindolylmaleimide I(1 μmol/L) compared with that in the control group (P < 0.05). With 3 μmol/L of bisindolylmaleimide I,the induced MCP-1 mRNA expression was reduced by 40% (P < 0.05). The LPA (10 μmol/L)-induced MCP-1 protein production was reduced by 78% with SP600125 (1 μmol/L) (P < 0.05). The 10 μmol/L LPA-induced MCP-1 mRNA expression was reduced by 40% with SP600125 (1 μmol/L) (P < 0.05). The activity of JNK was enhanced by LPA in HLF-1 while PKC inhibitors bisindolylmaleimide I (1 μmol/L) suppressed the activity of JNK which was induced by LPA(10 μmol/L). CONCLUSION: PKC and JNK mediated LPA-induced MCP-1 expression in HLF-1.

Key words: lysophosphatidic acid, protein kinase C, c-Jun N-terminal kinase, monocyte chemo-attractant protein-1, human lung fibroblasts

CLC Number:

R329.2

YIN Qi, XU Mingyan, FU Yucai, DENG Xiaoling. Protein kinase C and JN K mediate lysophosphatidic acid-induced monocyte chemo-attractant protein-1 expression in human fetal lung fibroblasts[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(1): 31-36.

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Protein kinase C and JN K mediate lysophosphatidic acid-induced monocyte chemo-attractant protein-1 expression in human fetal lung fibroblasts

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